Elevation of the adenylate pool in rat cardiomyocytes by S-adenosyl-L-methionine.
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Rapid resynthesis of the adenylate pool in cardiac myocytes is important for recovery of contractility and normal function of regulatory mechanisms in the heart. Adenosine and adenine are thought to be the most effective substrates for nucleotide synthesis, but the possibility of using other compounds has been studied very little in cardiomyocytes. In the present study, the effect of S-adenosyl-L-methionine (SAM) on the adenylate pool of isolated cardiomyocytes was investigated and compared to the effect of adenine and adenosine. Adult rat cardiomyocytes were isolated using the collagenase perfusion technique. The cells were incubated in the presence of adenine derivatives for 90 min followed by nucleotide determination by HPLC. The concentrations of adenine nucleotides expressed in nmol/mg of cell protein were initially 22.1 ± 1.4, 4.0 ± 0.3 and 0.70 ± 0.08 for ATP, ADP and AMP, respectively (n = 10, ±S.E.M.), and the total adenylate pool was 26.8 ± 1.6. In the presence of 1.25 mM SAM in the medium, the adenylate pool increased by 5.2 ± 0.4 nmol/mg of cell protein, but only if 1 mM ribose was additionally present in the medium. No changes were observed with SAM alone. A similar increase (by 4.9 ± 0.6 nmol/mg protein) was observed after incubation with 1.25 mM adenine plus 1 mM ribose, but no increase was observed if ribose was omitted. Adenosine at 0.1 or 1.25 mM concentrations also caused an increase in the adenylate pool (by 5.2 ± 1.0 and 5.2 ± 0.9 nmol/mg protein, respectively), which in contrast to the SAM or adenine was independent of the additional presence of ribose. Thus, S-adenosyl-L-methionine could be used as a precursor of the adenylate pool in cardiomyocytes, which is as efficient in increasing the adenylate pool after 90 min of incubation as adenosine or adenine. Nucleotide synthesis from SAM involves the formation of adenine as an intermediate with its subsequent incorporation by adenine phosphoribosyltransferase.
- 1. Achterberg, P.W. (1988) Adenine nucleotides, purine metabolism and myocardial function; in Myocardial Energy Metabolism (de Jong, J.W., ed.) pp. 45-52, Nijhof Publs, Dordrecht, Boston, Lancaster.
- 2. Kupriyanov, V.V., Lakomkin, V.L., Kapelko, V.I., Steinschneider, A.Y., Ruuge, E.K. & Saks, V.A. (1987) Dissociation of adenosine triphosphate levels and contractile function in isovolumetric hearts perfused with 2-deoxyglucose. J. Mol. Cell. Cardiol. 19, 729-740.
- 3. Smolenski, R.T., Simmonds, H.A. & Chambers, D.J. (1997) Exogenous adenosine supplied transiently during reperfusion ameliorates depressed endogenous adenosine production in the post-ischemic rat heart. J. Mol. Cell. Cardiol. 29, 333-346.
- 4. Berne, R.M. (1980) The role of adenosine in the regulation of coronary blood flow. Circ. Res. 47, 807-813.
- 5. Schrader, J., Baumann, G. & Gerlach, E. (1977) Adenosine as inhibitor of myocardial effects of catecholamines. Pflugers Arch. 372, 29-35.
- 6. Mills, D.C.B., MacFarlane, D.E., Lemmex, B.W.G. & Haslam, R.J. (1983) Receptors for nucleosides and nucleotides on blood platelets; in Regulatory Functions of Adenosine (Berne, R.M. et al., eds.) pp. 277-289. Nijhof Publs, The Hague, Boston, London.
- 7. Cronstein, B.N., Levin, R.I., Philips, M., Hirschhorn, R., Abramson, S.B. & Weissmann, G. (1992) Neutrophil adherence to endothelium is enhanced via adenosine A1 receptors and inhibited via adenosine A2 receptors. J. Immunol. 148, 2201-2206.
- 8. Brown, A.K., Raeside, D.L., Bowditch, J. & Dow, J.W. (1985) Metabolism and salvage of adenine and hypoxanthine by myocytes isolated from mature rat heart. Biochim. Biophys. Acta 845, 469-476.
- 9. Bowditch, J., Brown, A.K. & Dow, J.W. (1985) Accumulation and salvage of adenosine and inosine by isolated mature cardiac myocytes. Biochim. Biophys. Acta 844, 119-128.
- 10. Dow, J.W., Bowditch, J., Nigdikar, S.V. & Brown, A.K. (1987) Salvage mechanisms for regeneration of adenosine triphosphate in rat cardiac myocytes. Cardiovasc. Res. 21, 188- 196.
- 11. Zimmer, H.G. & Ibel, H. (1984) Ribose accelerates the repletion of the ATP pool during recovery from reversible ischemia of the rat myocardium. J. Mol. Cell. Cardiol. 16, 863- 866.
- 12. Montero, C., Smolenski, R.T., Duley, J.A. & Simmonds, H.A. (1990) S-adenosylmethionine increases erythrocyte ATP in vitro by a route independent of adenosine kinase. Biochem. Pharmacol. 40, 2617-2623.
- 13. Smolenski, R.T., Fabianowska-Majewska, K., Montero, C., Duley, J.A., Fairbanks, Marlewski, M. & Simmonds, H.A. (1992) A novel route of ATP synthesis. Biochem. Pharmacol. 43, 2053-2057.
- 14. Smolenski, R.T., Montero, C., Duley, J. & Simmonds, H.A. (1991) Effects of adenosine analogues on ATP concentration in human erythrocytes. Further evidence for a route independent of adenosine kinase. Biochem. Pharmacol. 42, 1767-1773.
- 15. Montero, C., Smolenski, R.T., Duley, J.A. & Simmonds, H.A. (1991) An alternative pathway of adenylate and ATP synthesis. Adv. Exp. Med. Biol. 309A, 285-288.
- 16. Smolenski, R.T., Schrader, J., de Groot, H. & Deussen, A. (1991) Oxygen partial pressure and free intracellular adenosine of isolated cardiomyocytes. Am. J. Physiol. 260, C708- C714.
- 17. Geisbuhler, T., Altschuld, R.A., Trewyn, R.W., Ansel, A.Z., Lamka, K. & Brierley, G.P. (1984) Adenine nucleotide metabolism and compartmentalization in isolated adult rat heart cells. Circ. Res. 54, 536-546.
- 18. Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265-275.
- 19. Smolenski, R.T., Lachno, D.R., Ledingham, S.J.M. & Yacoub, M.H. (1990) Determination of sixteen nucleotides, nucleosides and bases using high-performance liquid chromatography and its application to the study of purine metabolism in hearts for transplantation. J. Chromatogr. 527, 414-420.
- 20. Smolenski, R.T., Skladanowski, A.C., Swierczynski, J., Perko, M., Narkiewicz, M. & Zydowo, M.M. (1993) Changes of nucleotide content in human and rat heart during cardiac surgery and ischemia. Acta Biochim. Polon. 40, 531-538.
- 21. Maeda, N., Kon, K., Sekiya, M. & Shiga, T. (1986) Increase of ATP level in human erythrocytes induced by S-adenosyl-L-methionine. Biochem. Pharmacol. 35, 625-629.
- 22. Sahota, A., Webster, D.R., Potter, C.F., Simmonds, H.A., Rodgers, V.A. & Gibson, T. (1983) Methylthioadenosine phosphorylase activity in human erythrocytes. Clin. Chim. Acta 128, 283-290.
- 23. Hoffman, J.L. (1986) Chromatographic analysis of the chiral and covalent instability of S-adenosyl-L-methionine. Biochemistry 25, 4444-4449.
- 24. Revelle, L.K., d'Avignon, D.A., Reepmeyer, J.C. & Zerfing, R.C. (1995) Stability-indicating proton nuclear magnetic resonance spectroscopic method for determination of S-adenosyl-L-methionine in tablets. J. AOAC. Int. 78, 353-358.
- 25. Barber, J.R., Morimoto, B.H., Brunauer, L.S. & Clarke, S. (1986) Metabolism of S-adenosyl-L-methionine in intact human erythrocytes. Biochim. Biophys. Acta, 886, 361-372.
- 26. Stramentinoli, G., Pezzoli, C. & Kienle, M.G. (1978) Uptake of S-adenosyl-L-methionine by rabbit erythrocytes. Biochem. Pharmacol. 27, 1427-1430.
- 27. Avila, J.L. & Polegre, M.A. (1993) Uptake and metabolism of S-adenosyl-L-methionine by Leishmania mexicana and Leishmania braziliensis promastigotes. Mol. Biochem. Parasitol. 58, 123-134.
- 28. Gerbracht, U., Eigenbrodt, E., Simile, M.M., Pascale, R.M., Gaspa, L., Daino, L., Seddaiu, M.A., De Miglio, M.R., Nufris, A. & Feo, F. (1993) Effect of S-adenosyl-L-methionine on the development of preneoplastic foci and the activity of some carbohydrate metabolizing enzymes in the liver, during experimental hepatocarcinogenesis. Anticancer Res. 13, 1965-1972.
- 29. Lieber, C.S. (1994) Hepatic and metabolic effects of ethanol: Pathogenesis and prevention. Ann. Med. 26, 325-330.
- 30. Muriel, P., Suarez, O.R., Gonzalez, P. & Zuniga, L. (1994) Protective effect of S-adenosyl-L-methionine on liver damage induced by biliary obstruction in rats: A histological, ultrastructural and biochemical approach. J. Hepatol. 21, 95-102.
- 31. Alvaro, D., Gigliozzi, A., Piat, C., Carli, L., Bini, A., La Rosa, T., Furfaro, S. & Capocaccia, L. (1995) Effect of S-adenosyl-L-methionine on ethanol cholestasis and hepatotoxicity in isolated perfused rat liver. Dig. Dis. Sci. 40, 1592-1600.
- 32. Garcia Ruiz, C., Morales, A., Colell, A., Ballesta, A., Rodes, J., Kaplowitz, N. & Fernandez Checa, J.C. (1995) Feeding S-adenosyl-L-methionine attenuates both ethanol-induced depletion of mitochondrial glutathione and mitochondrial dysfunction in periportal and perivenous rat hepatocytes. Hepatology 21, 207-214.
- 33. Arch, J.R.S. & Newsholme, E.A. (1978) Activities and some properties of 5'-nucleotidase, adenosine kinase and adenosine deaminase in tissues from vertebrates and invertebrates in relation to the control of the concentration and the physiological role of adenosine. Biochem. J. 174, 965-977.
- 34. Smolenski, R.T. & Yacoub, M.H. (1996) Purine metabolism in human cardiomyocytes and endothelium implications for protection of the heart during cardiac surgery; in Purines and Myocardial Protection (Abd-Elfattah, A.S.A. et al., eds.) pp. 55-80, Kluwer Academic Publishers, Norwell.
- 35. Lieber, C.S. (1999) Role of S-adenosyl-L-methionine in the treatment of liver diseases. J. Hepatol. 30, 1155-1159.
- 36. Net, M., Garcia-Valdecasas, J.C., Deulofeu, R., Gonzalez, X., Palacin, J., Almenara, R., Valero, R., Lopez-Boado, M., Angas, J., Elena, M., Ballesta, A.M. & Visa, J. (1999) S-Adenosyl-L-methionine effect on hepatic allografts procured from non-heart-beating donor pigs. Transplant. Proc. 31, 1063-1064.
- 37. Benson, R., Crowell, B., Hill, B., Doonquah, K. & Charlton, C. (1993) The effects of L-dopa on the activity of methionine adenosyltransferase relevance to L-dopa therapy and tolerance. Neurochem. Res. 18, 325-330.
- 38. Simmonds, H.A., Van Acker, K.J., Cameron, J.S. & Snedden, W. (1976) The identification of 2,8-dihydroxyadenine, a new component of urinary stones. Biochem. J. 157, 485-487.
- 39. Cameron, J.S., Simmonds, H.A., Cadenhead, A. & Farebrother, D. (1977) Metabolism of intravenous adenine in the pig. Adv. Exp. Med. Biol. 76A, 196-205.
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